tbuckley/cabot-search huggingface dataset:
Abstract 3734: Neuroendocrine differentiation (ND) in sensitivity of neuroendocrine tumor (NET) cells to ONC201/TIC10 cancer therapeutic
Authors: Elizabeth C. Ding, Wafik S. El-Deiry
Journal: Cancer Research (2023)

Abstract Neuroendocrine tumors (NETs) harbor neuroendocrine differentiation (ND) with specific markers including protein gene product 9.5 (PGP9.5) and Chromogranin A (CgA). In prostate cancers (PC), ND is induced by BRN2/SOX2 transcription factors. NET-like cells with low or absent androgen receptor (AR) signaling cause hormone therapy resistance and poor prognosis in PC. Small cell lung carcinoma (SCLC), a high-grade NET, presents with metastasis early and has poor survival. ONC201/TIC10 is a small molecule inducer of TRAIL signaling in clinical trials. ONC201 antagonizes dopamine D2 or D3 receptors (DRD2/DRD3) and is an agonist of mitochondrial caseinolytic protease P (ClpP) resulting in activation of DR5/TRAIL-dependent apoptosis involving the integrated stress response (ISR). ONC201 is active in various malignancies including H3K27M-mutated glioma and NETs expressing high levels of DRD2. We hypothesized that altered BRN2/SOX2 may impact NET apoptosis by ONC201 through the ISR and TRAIL/DR5. We analyzed the expression of neuroendocrine markers PGP9.5, CgA, SOX2, and BRN2, as well as markers of TRAIL signaling pathway markers ATF4, DR5, ClpP, ClpX, and DRD2/DRD3 in PC and SCLC cell lines (N=6) ± treatment with ONC201. Specifically, we compared pre-treatment protein expression levels with the IC50. Our results reveal that DU145 (IC50=3.11μM), PC3 (IC50=3.02μM), and LNCaP (IC50=1.33μM) are ONC201 sensitive. H1417 SCLC expresses CgA, unlike PC3 and DU145. PGP9.5 is expressed in these lines. PGP9.5 is expressed in PC3, DU145, H1417, and H1048 but not in LNCaP and 22RV1. BRN2 is expressed in PC3, H1417, and H1048 but not DU145, LNCaP, or 22RV1. ClpX is expressed in all 6 lines but at lower levels in SCLC. ClpP is expressed in the 6 lines. DR5 is expressed at higher levels in PC3, DU145, LNCaP, and 22RV1 PC versus H1417 and H1048 SCLC. SOX2 is expressed at high levels in H1417 cells. These results are establishing the landscape of ND in PC and SCLC lines for further experimentation and testing of our hypothesis. To characterize the association of BRN2 dysregulation with ONC201 sensitivity, we are performing BRN2/SOX2 knockdown experiments using siRNA and evaluating effects towards ONC201 sensitivity. Our results provide insights into molecular mechanisms of ND in PC and SCLC sensitivity to ONC201. We are also currently working to overexpress BRN2 and SOX2 in cell lines to analyze the impact on neuroendocrine differentiation and ONC201 sensitivity. Citation Format: Elizabeth C. Ding, Wafik S. El-Deiry. Neuroendocrine differentiation (ND) in sensitivity of neuroendocrine tumor (NET) cells to ONC201/TIC10 cancer therapeutic. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3734.
Abstract 6111: Design of a small molecule screening assay to detect DNA trapping of PARP1/2
Authors: Kasia Zientara-Rytter, Veronique T. Baron, Junguk Park, Pavel Shashkin, Henry Zhu
Journal: Cancer Research (2023)

Abstract Poly(ADP-ribose) polymerase (PARP) inhibitors are currently used in the clinic for the treatment of tumors with a defective DNA damage response (DDR). When PARP1 or PARP2 binds damaged DNA, it adds poly(ADP-ribose) chains to its own backbone and to other DDR proteins, which recruits and activates them. PARylated PARP1/2 next detaches from the DNA so that the other PARylated proteins can initiate the repair process. It has been observed that some PARP inhibitors prevent PARP1/2 from dissociating the DNA. The continuous presence of PARP at the site of damage prevents repair and blocks replication, leading to cell death. Therefore, drugs that trap PARP1/2 to the DNA tend to be significantly more cytotoxic than other PARP inhibitors, which is highly desirable. This study describes the design and optimization of novel PARPtrap assays to specifically assess the ability of a drug to trap PARP onto DNA. The assay is based on principles of fluorescence polarization and uses fluorescently labeled DNA probes that are excited by polarized light and emit light with a degree of polarization that is proportional to the rate of molecular rotation. The free DNA probes that rotate fast have low fluorescence polarization (FP), but high FP when are bound to PARP1 or PARP2. When NAD+ is added, the PARylated enzymes detach from the probe, reducing FP levels. If a PARP inhibitor is added, the inhibitor’s trapping ability increases FP in a dose-dependent manner. Proof-of-principle titration of known PARP-trapping inhibitors (Talazoparib, AZD305, Olaparib and Veliparib) was performed to validate the assay. We observed that: i)The known relative trapping efficacies of Talazoparib, Olaparib and Veliparib were similar to known relative efficacies. ii)Talazoparib, Olaparib and Veliparib had similar trapping efficacy against PARP1 and PARP2, as measured by their EC50, whereas AZD305 was 1,000 times more efficient at trapping PARP1 than it was PARP2, demonstrating selectivity between PARP1 and PARP2. iii)AZD305 displayed as efficient DNA trapping activity toward PARP1 as best-in-class Talazoparib. In summary, we have designed an innovative PARPtrap assay designed for the high throughput screening of small molecule libraries to specifically identify or compare inhibitors that are capable of trapping PARP1 and/or PARP2 onto DNA. Citation Format: Kasia Zientara-Rytter, Veronique T. Baron, Junguk Park, Pavel Shashkin, Henry Zhu. Design of a small molecule screening assay to detect DNA trapping of PARP1/2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6111.
Abstract 1290: PTEN loss-mediated cell surface translocation of Annexin A2 promotes prostate cancer metastasis
Authors: Pankaj Chaudhary, Christianna Howard-Smith
Journal: Cancer Research (2023)

Abstract Introduction: Metastasis is a major cause of morbidity in prostate cancer patients. Despite substantial efforts to understand prostate cancer metastasis, the mechanisms involved in preparing the metastatic niche for colonizing the prostate cancer cells are still unknown. Methods/Materials: The normal prostate epithelial and cancer cells were purchased from American Type Culture Collection (Manassas, VA). Immunoblotting, immunofluorescence, flow cytometry, and plasmin generation assay were performed. Results: Our results demonstrated that Pten deletion or mutation caused significantly high phosphorylation of Annexin A2 (AnxA2) at tyrosine-23 in human and murine prostate cancer cell lines compared to the normal prostate epithelial cell lines. However, we did not observe any change in total AnxA2 protein level after Pten deletion/mutation in human and murine prostate cancer cells compared to normal prostate epithelial cells. Our flow cytometry data, immunofluorescence, and membrane wash fraction analysis suggest that AnxA2 expression was higher at the cell surface of Pten null/mutated prostate cancer cell lines compared to the normal prostate epithelial cells. Confocal microscopy and immunoprecipitation analysis revealed that AnxA2 interacts with Pten in normal prostate epithelial cells and inhibits AnxA2 phosphorylation at tyrosine 23. Pten deletion/mutation-mediated translocation of AnxA2 is dependent on the phosphorylation of tyrosine 23, and mutation of tyrosine 23 to a non-phosphomimetic variant inhibits the cell surface translocation process. The cell surface-translocated AnxA2 promotes the plasmin generation in prostate cancer cells, and an AnxA2 inhibitory hexapeptide can neutralize this activity. Conclusions: PTEN loss is the primary driving force for the translocation of AnxA2 to the cell surface and promotes plasmin generation. Citation Format: Pankaj Chaudhary, Christianna Howard-Smith. PTEN loss-mediated cell surface translocation of Annexin A2 promotes prostate cancer metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1290.
Abstract 1649: Chemically induced chaperone-mediated protein degradation (CHAMP) of KRAS(G12C)
Authors: Kevin P. Foley, Chenghao Ying, Yaya Wang, Yan Dai, Zhiyong Wang, Jinhua Li, Zimo Yang, Yuetong Sun, Hao Xin Zhou, Thomas L. Prince, Guoqiang Wang, Weiwen Ying
Journal: Cancer Research (2023)

Abstract Covalent KRAS(G12C) inhibitors have recently shown promising efficacy in the clinic. However, the rapid development of drug resistance compromises their long-term benefits, indicating the need for additional approaches to treat KRAS-mutated cancers. Targeted protein degradation represents one such potential approach. We have therefore explored applying Chaperone-mediated Protein Degradation (CHAMP) technology to degrade KRAS(G12C) protein. CHAMPs are heterobifunctional small molecules that mediate protein degradation by inducing proximity between a target protein and the HSP90 chaperone complex, resulting in target ubiquitination and degradation by the proteasome. Due HSP90 being highly activated in cancer cells, CHAMPs also preferentially accumulate in tumors relative to normal tissues, resulting in an improved therapeutic index relative to typical inhibitors. In order to chemically induce KRAS(G12C) degradation, CHAMP compounds were synthesized by covalently coupling KRAS(G12C)- and HSP90-binding moieties through a short linker. CHAMP treatment of KRAS(G12C)-mutated cell lines resulted in formation of a KRAS(G12C)-CHAMP-HSP90 ternary complex and subsequent proteasome-dependent KRAS(G12C) degradation and inhibition of cell proliferation. In contrast, combination treatment with KRAS(G12C) and HSP90 inhibitors did not result in substantial KRAS(G12C) degradation. Importantly, CHAMPs retained potency in cell line models of resistance to covalent KRAS(G12C) inhibitors. Further, in mouse xenograft models, CHAMPs displayed prolonged pharmacokinetics in tumors relative to plasma and normal tissues and strongly inhibited tumor growth at tolerated doses. Based on these findings, KRAS(G12C)-degrading CHAMPs represents a promising approach to the treatment of KRAS(G12C)-mutated cancers. Citation Format: Kevin P. Foley, Chenghao Ying, Yaya Wang, Yan Dai, Zhiyong Wang, Jinhua Li, Zimo Yang, Yuetong Sun, Hao Xin Zhou, Thomas L. Prince, Guoqiang Wang, Weiwen Ying. Chemically induced chaperone-mediated protein degradation (CHAMP) of KRAS(G12C) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1649.
Abstract 4699: Pan-cancer single-cell analysis reveals the interactions of <i>SPP1+MMP12+</i> macrophages with metastasis-initiating cells that drives metastasis
Authors: Baifeng Zhang, Dora Lai-wan Kwong, Xin-Yuan Guan
Journal: Cancer Research (2023)

Abstract Advanced understanding of cancer immunology has contributed to a significant decline in cancer deaths in the past few decades. However, there are still vast majority of patients with no effective response for current immunotherapy drugs because of some innate immune cells, particularly macrophages and neutrophils, promoting all steps of malignant progression to metastasis. Therefore, the development of new strategies against these pro-metastatic non-tumor components in the tumor microenvironment is in need. Tumor-associated macrophage (TAMs), as the critical component of tumor microenvironment (TME), display diverse and even opposing function phenotypes, but without systematic investigation on their ubiquitous characteristics across different cancer types. Here, we perform pan-cancer analysis on 92 samples across 6 solid cancer types to profile the TME of normal, primary tumor, metastatic lymph node and distant metastasis at single-cell resolution, illustrating the commonalities/plasticity of heterogenous TAMs. We uncover 21 distinct subtypes of TAMs with varying abundance and biological function in different tissue microenvironment, providing evidence for the plasticity of TAMs. In particular, the enrichment of SPP1+MMP12+ macrophages in the TME of lymph node metastasis, likely interacting with metastasis-initiating cells (MICs), is associated with promoting malignant progression to metastasis and survival stratifications. Multi-color immunofluorescence (mIF) staining for relevant markers is being performed in mouse models and clinical samples for comprehensive validation. Our study comprehensively describes the characteristics and dynamics of TAMs and highlights their heterogeneity and plasticity depending on their surrounding microenvironment. Citation Format: Baifeng Zhang, Dora Lai-wan Kwong, Xin-Yuan Guan. Pan-cancer single-cell analysis reveals the interactions of SPP1+MMP12+ macrophages with metastasis-initiating cells that drives metastasis. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4699.
Abstract 4904: Glutamine antagonist DRP-104 enhances anti-tumor responses in <i>KEAP1</i> mutant lung cancer
Authors: Ray Pillai, Sarah LeBoeuf, Ali Rashidfarrokhi, Shih Ming Huang, Triantafyllia Karakousi, Anastasia-Maria Zavitsanou, Warren Wu, Volkan Sayin, Robert Wild, Sergei Koralov, Thales Papagiannakopoulos
Journal: Cancer Research (2023)

Abstract KEAP1 is mutated in approximately 20% of non-small cell lung cancer (NSCLC) and is associated with poor response rates to checkpoint blockade. Given the therapeutic challenge presented by KEAP1 mutant tumors and the great potential held by immunotherapeutic approaches, there is an urgent need to identify the mechanisms that mediate immune evasion in patients with KEAP1 mutant lung adenocarcinoma. We previously demonstrated that metabolic rewiring in KEAP1 mutant tumors leads to increased dependency on glutamine as a substrate for multiple anabolic pathways. DRP-104 (Sirpiglenastat) is a novel glutamine antagonist which inhibits all glutamine consuming reactions. Using pre-clinical mouse models and patient derived xenograft models we demonstrate that DRP-104 robustly impairs the growth of KEAP1 mutant tumors. Furthermore, using an immunocompetent orthotopic lung adenocarcinoma mouse model we find that treatment with DRP-104 increases survival of Keap1 mutant tumor bearing mice treated with anti-PD1 therapy. We have dissected the mechanism by which DRP-104 impacts KEAP1 mutant tumors and the surrounding immune microenvironment. Notably, DRP-104 impairs nucleotide synthesis in KEAP1 mutant tumors and thereby inhibits cancer cell growth. In addition, DRP-104 reduces exhausted T cell and T regulatory populations within the tumor microenvironment as well as increases effector T cell function. Overall, we find that DRP-104 impairs the growth of KEAP1 mutant tumors through both targeting of tumor intrinsic metabolic vulnerabilities and through tumor extrinsic immunostimulatory mechanisms such as promoting the expansion of functional anti-tumor T cell populations. This data provides additional rationale for the ongoing phase I/II clinical trial (NCT04471415) using DRP-104 in NSCLC with mutations in KEAP1 as well as for combining DRP-104 with checkpoint blockade. Citation Format: Ray Pillai, Sarah LeBoeuf, Ali Rashidfarrokhi, Shih Ming Huang, Triantafyllia Karakousi, Anastasia-Maria Zavitsanou, Warren Wu, Volkan Sayin, Robert Wild, Sergei Koralov, Thales Papagiannakopoulos. Glutamine antagonist DRP-104 enhances anti-tumor responses in KEAP1 mutant lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4904.
Abstract 2804: 3D printing of perfusable alveolar model for efficacy evaluation of polyisoprenylated cysteinyl amide inhibitors on lung cancer cells
Authors: Diana Isabel Romero, Bing Ren, Christopher Leon, Ite A. Offringa, Nazarius S. Lamango, Yong Huang
Journal: Cancer Research (2023)

Abstract Lung cancer is the leading cause of cancer death in the United States, and Black Americans are being disproportionality affected. Lung adenocarcinoma (LUAD) is the most common histological lung cancer subtype among all lung cancer patients, including black patients. There is still no available physiologically relevant three-dimensional (3D) in vitro model that can be used to study the treatment efficacy on LUAD cells, in particular, those from Black patients. In this study, we aim to develop a 3D alveolar model with a perfusable epithelialized channel using 3D biofabrication technologies. We will apply the model to test their efficacies of polyisoprenylated cysteinyl amide inhibitors (PCAIs) and other drugs on LUAD. First, a perfusable channel is constructed within a gelatin composite extracellular matrix (ECM) using a sacrificial material-assisted 3D embedded printing technology. Next, the channel is seeded with KRAS-mutated NCI-H23 LUAD (derived from a Black patient) or hAEC-C7 (untransformed, immortalized human alveolar epithelial cells) respectively, and exposed to drugs to test the anti-cancer efficacy. In particular, PCAIs, which are potential therapeutics targeting activated KRAS, are investigated for their cytotoxicity on H23 LUAD and hAEC-C7 cells for comparison. Compared to two-dimensional (2D) models, the proposed 3D alveolar model can offer a more physiologically relevant 3D cell-to-ECM environment to test LUAD therapeutics. The use of H23 cells and, in the future, other LUAD cells from Black patients ensures that any developed drugs constitute valid treatment options for black patients with RAS-mutated pathways, thus minimizing health disparities. Citation Format: Diana Isabel Romero, Bing Ren, Christopher Leon, Ite A. Offringa, Nazarius S. Lamango, Yong Huang. 3D printing of perfusable alveolar model for efficacy evaluation of polyisoprenylated cysteinyl amide inhibitors on lung cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2804.
Abstract 1793: Single-cell RNA analysis reveal effector-like CD8+ CAR-T cell subpopulations associated with response in lymphoma patients
Authors: Tina Sarén, Mohanraj Ramachandran, Gustav Gammelgård, Tanja Lövgren, Kristina Wikström, Jamileh Hashemi, Eva Freyhult, Claudio Mirabello, Åsa K Björklund, Håkan Ahlström, Rose-Marie Amini, Hans Hagberg, Angelica Loskog, Gunilla Enblad, Magnus Essand
Journal: Cancer Research (2023)

Abstract Although CD19-targeting CAR T cell therapy has shown remarkable success against B cell malignancies there is still room for improvement. To improve long-term responses amongst lymphoma patients, mechanisms underlying initial response and tumor relapses must be addressed. Recent studies have highlighted the importance of the quality of the starting cell material for CAR T cell production and the composition of the CAR T cell infusion product for clinical response. In our study, we have evaluated the clinical response to third generation anti-CD19 CAR T cell therapy in 23 patients with B cell lymphoma. Further, we evaluated the CAR T cell infusion products using targeted single-cell RNA sequencing and multi-color flow cytometry to find potential T cell subsets associating with clinical response. Patients were divided into responders (CR + PR) and non-responders (SD + PD) at the 1-month follow-up after treatment. The CAR T cell products of both responding and non-responding patients were dominated by CD8+ CAR T cells. When comparing cells from the two patient groups we found that the presence of effector-like CD8+ CAR T cells with a high cytotoxic and cytokine secretion and low dysfunction gene expression profile were associated with clinical response. Furthermore, CD8+ CAR T cells of responders displayed a higher polyfunctional gene expression profile, which has previously been shown to associate with clinical response. In contrast, CD8+ CAR T cells from the infusion products of non-responders displayed an elevated dysfunction gene expression profile. Our study reveals that single-cell RNA expression analysis of individual CAR T cell infusion product can identify subsets of effector cells correlating with clinical response. This can be used as a guidance in the CAR T cell production processes, to obtain CAR T cells with the desired characteristics. Citation Format: Tina Sarén, Mohanraj Ramachandran, Gustav Gammelgård, Tanja Lövgren, Kristina Wikström, Jamileh Hashemi, Eva Freyhult, Claudio Mirabello, Åsa K Björklund, Håkan Ahlström, Rose-Marie Amini, Hans Hagberg, Angelica Loskog, Gunilla Enblad, Magnus Essand. Single-cell RNA analysis reveal effector-like CD8+ CAR-T cell subpopulations associated with response in lymphoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1793.
Abstract 1602: Co-targeting mitochondria and nucleotide metabolism in T315I-BCR-ABL myeloid leukemia for therapy
Authors: Chang-Yu Huang, Yin-Hsuan Chung, Sheng-Yang Wu, Hsin-Yang Wang, Chih-Yu Lin, Tsung-Jung Yang, Jim-Ming Fang, Yu-Ju Chen, Chun-Mei Hu, Zee-Fen Chang
Journal: Cancer Research (2023)

Abstract T315I mutation of Bcr-Abl in chronic myeloid leukemia (CML) leads to therapeutic resistance. It is known that Bcr-Abl transformation causes ROS-induced DNA damage, which can be exploited for anti-nucleotide treatment. We developed a small compound, JMF4073, which inhibits pyrimidylate kinases. Intriguingly, treatment of JMF4073 selectively eliminated WT-, but not T315I-, Bcr-Abl-transformed cells. We found that the higher level of cellular glutathione(GSH) in T315I-Bcr-Abl cells confers replication fork progression and sustains dTTP pool, leading to JMF4073 insusceptibility. Blocking mitochondrial pyruvate carrier (MPC) by UK-5099 reduced GSH and induced DNA replication stress in T315I-Bcr-Abl cells, thus increasing JMF4073 sensitivity in vitro and in vivo. We also observed the increases in glutamine flux to GSH production in T315I-Bcr-Abl cells, and reductive carboxylation of glutamate metabolism, together with mitochondrial dysfunction. The proteomic assay revealed the deficiencies in respiration complex I, II, and IV proteins in T315I-Bcr-Abl cells. Most interestingly, calcineurin inhibition remarkably elevated mitochondrial oxidative stress to cause cytotoxicity specifically in T315I-Bcr-Abl cells. Our results demonstrate the metabolic shift by T315I mutation of Bcr-Abl CML and provide the therapeutic options by co-targeting mitochondria and pyrimidylate kinases. Citation Format: Chang-Yu Huang, Yin-Hsuan Chung, Sheng-Yang Wu, Hsin-Yang Wang, Chih-Yu Lin, Tsung-Jung Yang, Jim-Ming Fang, Yu-Ju Chen, Chun-Mei Hu, Zee-Fen Chang. Co-targeting mitochondria and nucleotide metabolism in T315I-BCR-ABL myeloid leukemia for therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1602.